Isolation and growth of the grasshopper pathogen, Entomophthora grylli

Abstract Entomophthora grylli was isolated and grown in pure culture in protoplast form. Cultures were obtained by germinating conidia collected from naturally infected adult Carolina grasshoppers, Dissosteira carolina , in Grace's tissue culture medium supplemented with 5% fetal bovine serum and an aqueous extract of grasshopper tissue. Grasshopper extract was not necessary for subsequent growth and subculture of the protoplasts. Healthy adult grasshoppers could be reinfected with the fungus by injection of a protoplast suspension.